Flow Cytometry to Evaluate Acrosome-Reacted Sperm

Miyazaki, Ryoichiro; Fukuda, Masaru; Takeuchi, Hiroyuki; Itoh, Shigeru; Takada, Makiko

doi:10.3109/01485019008987613

Cited by 24 publications

(7 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The superparamagnetic microbeads, which are about 50 nm in diameter, are too small to be detected by optical methods or by changing the scatter properties of spermatozoa in the flow cytometer (Miltenyi et al, 1990). As a positive control, the binding of FITC‐conjugated Pisum sativum agglutinin to the acrosome was employed according to the method described by Miyazaki et al (1990) for setting the bitmap on a dot plot. The negative controls represented spermatozoa that were treated with buffer containing immunoglobulin G 1 only (Glander and Schaller, 1999).…”

Section: Methodsmentioning

confidence: 99%

Deterioration of Plasma Membrane Is Associated With Activated Caspases in Human Spermatozoa

Paasch¹,

Grunewald²,

Fitzl

et al. 2003

Journal of Andrology

130

View full text Add to dashboard Cite

Spermatozoa with deteriorated plasma membranes can be separated by magnetic-activated cell sorting (MACS) after binding superparamagnetic annexin V-conjugated microbeads (ANMBs) to membrane phosphatidylserine (PS). Semen samples from 15 donors and 25 infertile patients were divided into 2 spermatozoal fractions by annexin V-MACS. Activated caspases (aCPs), which mediate degradations of cell quality, were determined by CaspaTag in the 2 subpopulations. Spermatozoa from donors showed lower levels of bound annexin V (3.6% Ϯ 0.5% vs 11.9% Ϯ 1.1%; P Ͻ .01) and aCPs (21.8% Ϯ 2.6% vs 43.2% Ϯ 2.1%; P Ͻ .01) than did spermatozoa from infertile patients. MACS resulted in a decrease of spermatozoa with aCPs from 21.8% Ϯ 2.6% (before separation) to 9.2% Ϯ 1.4%(in the ANMB-negative fraction) in donors and from 43.2% Ϯ 2.1% to 18.8% Ϯ 2.6% in infertile patients (mean Ϯ SEM; P Ͻ .01). Separation effects of the MACS technique were confirmed with flow cytometry using anti-annexin V antibodies and with electron microscopy. ANMB-MACS removes spermatozoa with PS-bound annexin V and produces a higher quality spermatozoal fraction. Spermatozoa with a deteriorated membrane are characterized by an increase in aCPs. A higher percentage of spermatozoa with ANMBs bound to PS and with aCPs were found in infertile patients.

show abstract

Section: Methodsmentioning

confidence: 99%

Deterioration of Plasma Membrane Is Associated With Activated Caspases in Human Spermatozoa

Paasch¹,

Grunewald²,

Fitzl

et al. 2003

Journal of Andrology

130

View full text Add to dashboard Cite

show abstract

“…Flow cytometry has been extensively used in veterinary and human andrology since its introduction in the 70s of the past century; initially focused on the analysis of sperm DNA (Dean, Pinkel, & Mendelsohn, ; Meistrich, Lake, Steinmetz, & Gledhill, ; Pinkel et al., ; Van Dilla et al., ). Later on, the applications of flow cytometry expanded to the study of the integrity of the membrane, acrosome and mitochondrial function (Engh, Clausen, & Purvis, ; Evenson, Darzynkiewicz, & Melamed, ; Graham, Kunze, & Hammerstedt, ; Miyazaki, Fukuda, Takeuchi, Itoh, & Takada, ; Ronot & Auger, ). The possibility to detect differences in DNA content allowed the development of the sexed semen market in the bovine industry (Johnson, Flook, & Look, ; Morrell, Keeler, Noakes, Mackenzie, & Dresser, ; Sharpe & Evans, ; Tubman, Brink, Suh, & Seidel, ; Umezu, Hiradate, Numabe, Hara, & Tanemura, ; Vazquez et al., ).…”

Section: Introductionmentioning

confidence: 99%

Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Peña

Rodriguez

Gil

et al. 2018

Reprod Domestic Animals

View full text Add to dashboard Cite

Contents Flow cytometry is increasingly used in research and also in clinical andrology. Recent developments in instrumentation, availability of probes and bioinformatics expand the possibilities of flow cytometry well beyond the classical two parametric analyses in use. In this paper, an overview of recent developments in flow cytometry will be presented under the perspective of the authors; aspects such a multicolor assays and computational cytometry will be discussed as well.

show abstract

“…Recently, Miyazaki et al (1990) have monitored the AR with flow cytometry. This method is more rapid and allows the evaluation of 10,000 spemiatozoa in a few minutes (Haynes, 1988).…”

Section: Introductionmentioning

confidence: 99%

Flow cytometric evaluation of the acrosome reaction of human spermatozoa: a new method using a photoactivated supravitaI stain

Henley

Roberts

1994

Int J of Andrology

View full text Add to dashboard Cite

A flow cytometric assay using a double-stain method for the measurement of the acrosome reaction of human spermatozoa is described. The use of a stable photoactivated stain, ethidium monoazide, allowed evaluation of the viability of spermatozoa. This stain was more stable in fixed samples than propidium iodide, which is not bound covalently to DNA and is therefore removed readily during the washing procedure. The permeabilized acrosome was labelled with Pisum sativum agglutinin conjugated with fluoroisothiocyanate. Since this lectin binds to the acrosome and acrosomal contents, a decrease in the fluorescence intensity allows the cytometric evaluation of the acrosome reaction. Microscopic analysis and flow cytometric analysis were well correlated and cell sorting was performed to ensure the homogeneity of each different subpopulation encountered.

show abstract

Flow Cytometry to Evaluate Acrosome-Reacted Sperm

Cited by 24 publications

References 30 publications

Deterioration of Plasma Membrane Is Associated With Activated Caspases in Human Spermatozoa

Deterioration of Plasma Membrane Is Associated With Activated Caspases in Human Spermatozoa

Flow cytometry analysis of spermatozoa: Is it time for flow spermetry?

Flow cytometric evaluation of the acrosome reaction of human spermatozoa: a new method using a photoactivated supravitaI stain

Contact Info

Product

Resources

About